QEEN II: 2ND QUANTIFYING EXPOSURE TO ENGINEERED NANOMATERIALS FROM - PowerPoint PPT Presentation

QEEN II: 2ND QUANTIFYING EXPOSURE TO ENGINEERED NANOMATERIALS FROM MANUFACTURED PRODUCTS WORKSHOP SESSION B. CONSUMER EXPOSURE: FOOD, FOOD CONTACT, AND PERSONAL CARE PRODUCTS Cytotoxicity, hemotoxicity, and in-vivo toxicity of surface modified PLGA nanoparticles Cristina M. Sabliov, LSU Alumni Professor Washington DC, October 8-10

Engineered nanoparticles Chou, L. Y. T, K. Ming and W. C. W. Chan. 2011. Chem. Soc. Rev. 40, 233-245.

• Intentional exposure • Fortified foods • Unintentional exposure • Packaging • Agrochemical nanodelivery systems Exposure Yu Yang, Paul Westerhoff. 2014. Nanomaterial Impacts on Cell Biology and Medicine , pp 1 – 17. Part of the Advances in Experimental Medicine and Biology book series (volume 811).

Picture drawn by Thanida Chuacharoen Cockburn et al., 2012 Particle ADME profile

Particle Biotransformation D.Shevlin, N.O'Brien, E.Cummins. 2018. Science of the Total environment. April, 2018.

PLGA 33K pH 6.5 0 hrs 6 hrs 24 hrs PLGA-Chi 20 K & 33K pH 6.5 pH stability (pH=6.5) Murugeshu, A., C. E. Astete, C. Leonardi, and C. M. Sabliov. 2011. Nanomedicine Vol. 6, No. 9, Pages 1513-1528.

Cytotoxicity Trif, M, P. E. Florian, A. Roseanu, M. Moisei, O. Craciunescu, C. E. Astete and C. M. Sablio liov. 2015. Journal of Biomedical Materials Research Part A . 103(1):3599-3611 Alqahtani, S., L. Simon, C. E. Astete, A. Alayoubi, P. W. Sylvester, S. Nazzal, Y. Shen, Z. Xu, A. Kaddoumi, C. M. Sabliov. 2015 . Journal of Colloid and Interface Science. 445: 243-251.

Red blood cells with Red blood cells with 2mg/ml NPs 3mg/ml NPs Untreated red blood cells Red blood cells with Red blood cells with 10mg/ml NPs 50mg/ml NPs Nanoparticles RBC Association (SEM Image)

94.48 90 NP Association Hemolysis 75 73.46 60 49.71 45 Hemolysis 37.21 Percentage 35.43 30 29.92 23.32 22.45 15 18.56 17.65 17.78 17.26 17.41 16.74 16.22 13.95 0.22 0 Negative 2 3 5 8 10 20 50 Positive Control Control Nanoparticle Concentration (mg/ml) Hemotoxicity

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PLGA PLGA-Chi (a) (d) PLGA 7 days (% daily dose) PLGA/Chi 7 days (% daily dose) PLGA 7 days (mg) PLGA/Chi 7 days (mg) 7 days 50 1.40 50 1.40 1.20 1.20 mg NP after 7 days mg Np after 7 days 40 40 % daily dose 1.00 1.00 % daily doe 30 30 0.80 0.80 0.60 0.60 20 20 0.40 0.40 10 10 0.20 0.20 0 0.00 0 0.00 Int Liver Kidney Spleen Lung Heart Brain Int Liver Kidney Spleen Heart Lung Brain Organ Organ (e) (b) 14 days PLGA/Chi 14 days (% daily dose) PLGA 14 days (% daily dose) 50 1.40 PLGA/Chi 14 days (mg) PLGA 14 days (mg) 50 1.40 1.20 mg Np after 14 days 40 1.20 mg NP after 14 days 40 1.00 % daily dose % daily dose 1.00 30 0.80 30 0.80 0.60 20 0.60 20 0.40 0.40 10 10 0.20 0.20 0 0.00 0 0.00 Int Liver Kidney Spleen Lung Heart Brain Int Liver Kidney Spleen Heart Lung Brain Organ Organ 21 days (c) PLGA 21 days (% daily dose) (f) PLGA/Chi 21 days (% daily dose) PLGA 21 days (mg) PLGA/Chi 21 days (mg) 50 1.40 50 1.40 mg NP after 21 days 1.20 1.20 40 40 % daily dose mg NP after 21 days 1.00 % daily dose 1.00 30 30 0.80 0.80 0.60 0.60 20 20 0.40 0.40 10 10 0.20 0.20 0 0.00 0 0.00 Int Liver Kidney Spleen Lung Heart Brain Int Liver Kidney Spleen Heart Lung Brain Organ Organ In-vivo biodistribution Navarro, S. T. Morgan, C. E. Astete, R. Stout, D. Coulon, P. Mottramand C. M. Sablio liov. 2016.. Nanomedicine. 11(13):1653-1669. (doi: 10.2217/nnm-2016-0022)

Control PLGA NPs Liver 14 days PLGA treatment shows increased numbers of Liver lymphocytes, plasma cells (black arrow) and occasional mast cells (white arrow). Spleen No significant lesions have been observed in spleen. Intestine villi show the presence of Intestinal scattered lymphocytes (blue arrow), plasma cells (red arrow) and villi histiocytic cells (white arrow) in the lamina propria. Arrows indicate scattered mitotic figures (black arrows). Increased Intestinal hyperplasia of intestinal crypt cells crypts indicated increased intestinal mucosal epithelial cell turnover.

• PLGA NPs were not cytotoxic, except at high concentration (>5 mg/ml) • Interaction between PLGA nanoparticles and RBCs was concentration dependent • PLGA nanoparticles associated with RBC membrane and had no hemotoxic effect at concentrations lower than 5 mg/ml (>5% increased over negative control), and was dependent on the surfactant • Minimal inflammatory changes were observed in the hepatic portal regions of the liver and the lamina propria of the small intestine in PLGA and PLGA-Chi dosed rats versus controls • The intestine had mild crypt hyperplasia, indicative of an increased intestinal mucosal epithelial turnover rate • No significant histologic lesions were seen in the lung, kidney, spleen, or brain in PLGA or PLGA-Chi dosed rats versus controls Conclusions

• Properties of nanomaterial still important and should be documented • Nanomaterial biotransformation is key to in-vivo behavior • Toxicity work is mainly focused on inorganic , metal and metal oxides nanoparticles • Interest is growing in biodegradable particles • Methods of detection of biodegradable particles in complex media need to be developed • Toxicity testing needed for long term , small concentration exposure is needed THANK YOU!